Selection of ceramic fluorapatite-binding peptides from a phage display combinatorial peptide library: optimum affinity tags for fluorapatite chromatography.

摘要: Peptide affinity tags have become efficient tools for the purification of recombinant proteins from biological mixtures. The most commonly used ligands in this type chromatography are immobilized metal ions, proteins, antibodies, and complementary peptides. However, major bottlenecks technique still related to ligands, including their low stability, difficulties immobilization, leakage into final products. A model approach is presented here overcome these by utilizing macroporous ceramic fluorapatite (CFA) as stationary phase CFA-specific short peptides tags. CFA chromatographic materials act both support matrix ligand. Peptides that bind with were identified a randomized phage display heptapeptide library. total five rounds selection performed. common N-terminal sequence was found two selected peptides: F4-2 (KPRSMLH) F5-4 (KPRSVSG). peptide F5-4, displayed more than 40% phages analyzed fifth round selection, subjected further studies. Selectivity chemical composition morphology assured adsorption dissociation constant, obtained F5-4/CFA isotherm, micromolar range, maximum capacity 39.4 nmol/mg. behavior characterized on different buffers. Preferential specific retention properties suggest possible application phage-derived tag enhancing selective recovery proteins. Copyright © 2013 John Wiley & Sons, Ltd.